Multifunctional Magnetic Nanoparticles: Design,

Synthesis, and Biomedical Applications

JINHAO GAO,† HONGWEI GU,‡ AND BING XU*,†,§
§
Department of Chemistry, Brandeis University, 415 South Street, Waltham,
Massachusetts 02454, †Department of Chemistry, The Hong Kong University of
Science and Technology, Clear Water Bay, Hong Kong, China, ‡College of
Chemistry, Chemical Engineering and Materials Science, Soochow University,
Suzhou, Jiangsu 215123, China

RECEIVED ON JANUARY 8, 2009

CON SPECTUS

T he combination of nanotechnology and molecular biology has developed into an emerging research area: nano-
biotechnology. Magnetic nanoparticles are well-established nanomaterials that offer controlled size, ability to be
manipulated externally, and enhancement of contrast in magnetic resonance imaging (MRI). As a result, these nano-
particles could have many applications in biology and medicine, including protein purification, drug delivery, and med-
ical imaging.
Because of the potential benefits of multimodal functionality in biomedical applications, researchers would like to
design and fabricate multifunctional magnetic nanoparticles. Currently, there are two strategies to fabricate mag-
netic nanoparticle-based multifunctional nanostructures. The first, molecular functionalization, involves attaching anti-
bodies, proteins, and dyes to the magnetic nanoparticles. The other method integrates the magnetic nanoparticles with
other functional nanocomponents, such as quantum dots (QDs) or metallic nanoparticles. Because they can exhibit sev-
eral features synergistically and deliver more than one function simultaneously, such multifunctional magnetic nano-
particles could have unique advantages in biomedical applications.
In this Account, we review examples of the design and biomedical application of multifunctional magnetic nano-
particles. After their conjugation with proper ligands, antibodies, or proteins, the biofunctional magnetic nanopar-
ticles exhibit highly selective binding. These results indicate that such nanoparticles could be applied to biological
medical problems such as protein purification, bacterial detection, and toxin decorporation. The hybrid nanostruc-
tures, which combine magnetic nanoparticles with other nanocomponents, exhibit paramagnetism alongside features
such as fluorescence or enhanced optical contrast. Such structures could provide a platform for enhanced medical imag-
ing and controlled drug delivery. We expect that the combination of unique structural characteristics and integrated
functions of multicomponent magnetic nanoparticles will attract increasing research interest and could lead to new
opportunities in nanomedicine.

Published on the Web 05/28/2009 www.pubs.acs.org/acr Vol. 42, No. 8 August 2009 1097-1107 ACCOUNTS OF CHEMICAL RESEARCH 1097
10.1021/ar9000026 CCC: $71.50 © 2009 American Chemical Society
Multifunctional Magnetic Nanoparticles Gao et al.
FIGURE 1. The scheme illustrates two strategies to fabricate multifunctional magnetic nanoparticles and their potential applications.
1. Introduction
The integration of nanotechnology with molecular biology
and medicine has resulted in active developments of a new
emerging research area, nanobiotechnology,1 which offers
exciting opportunities for discovering new materials, pro-
cesses, and phenomena. Nanoscale magnetic materials
have their own advantages that provide many exciting
opportunities in biomedical applications. First, they deliver
controllable sizes ranging from a few up to tens of nano-
meters, so their optimization of sizes and properties easily
matches with the interest of study. Second, the nanopar-
ticles can be manipulated by an external magnetic force.
This “action at a distance” provides tremendous advantages
for many applications. Third, magnetic nanoparticles play
an important role as MRI contrast enhancement agents
because the signal of magnetic moment of a proton around
magnetic nanoparticles can be captured by resonant
absorption. Recently, techniques and procedures for pro-
ducing monodispersed and size-controllable magnetic
nanoparticles (e.g., FePt, Fe3O4, and γ-Fe2O3) have advanced
considerably,2,3 which leads to very active explorations of
the applications of magnetic nanoparticles, including
biomedicine.
In this Account, we describe some potentially useful
designs and applications of multifunctional magnetic nano-
particles for biomedicine, which have been attracting
increased research effort because of their easily accessible
multimodality. Generally, there are two kinds of strategies
to fabricate magnetic nanoparticle-based multifunctional
nanostructures (Figure 1). The first strategy is molecular
functionalization. Biofunctional molecules (e.g., antibod-
ies, ligands, or receptors) coat the magnetic nanoparticles
and make them interact with a biological entity with high
affinity, thus providing a controllable means of “tagging”.
After molecular functionalization, the biofunctional mag-
netic nanoparticles confer high selectivity and sensitivity for
1098 ACCOUNTS OF CHEMICAL RESEARCH 1097-1107 August 2009
many biological applications. The second one is to com-
bine magnetic nanomaterials and other functional nano-
structures by sequential growth or coating, which produces
a single entity conferring multiple functions in nanoscale.
For example, using magnetic nanoparticles as seeds, the
growth of semiconducting chalcogenides produces
core-shell or heterodimer nanostructures with both mag-
netic and fluorescent properties, which lead to the demon-
stration of intracellular manipulation of nanoparticles and
a promising candidate for dual-functional molecular imag-
ing (i.e., combining MRI and fluorescence imaging). The
integration of magnetic and metallic nanoparticles forms
heterodimer structures that offer two distinct surfaces and
properties to allow different kinds of functional molecules
to attach onto the specific parts of the heterodimers, which
may bind to multiple receptors or act as agents for multi-
modality imaging. The encapsulation of a potential anti-
cancer drug by iron oxide nanoshells results in the
yolk-shell nanostructures, which promise novel nanode-
vices for controlled drug delivery.
2. Molecular Functionalization of Magnetic
Nanoparticles
Magnetic nanoparticles can take the advantage of specific
binding to detect or purify the biological entities after being
modified by biomolecules. Because of their unique property,
response to a magnetic field, biofunctional magnetic nano-
particles exhibit two features, specificity and magnetism. Most
surface modification strategies, either polymer coating or
chemical ligand exchange, are based on or derived from self-
assembled monolayers (SAMs).4,5 The following sections dis-
cuss the attachment of functional molecules onto magnetic
nanoparticles for several applications.
2.1. Specific Binding and Targeting. The interactions of
biomolecular pairs with specific high affinity exist prevailingly
in nature. If one of the biomolecular entities conjugates with
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 Multifunctional Magnetic Nanoparticles Gao et al.

the magnetic nanoparticles, the resulting biofunctional mag-

netic nanoparticles can specifically bind to the other biomo-
lecular entity. Then, the most direct consequence is that the
external magnetic force can control the location of the bio-
logical entity. Based on this concept, several applications using
biofunctional magnetic nanoparticles, such as pathogen detec-
tion, protein purification, and toxin decorporation, have been
demonstrated in research laboratories.
2.1.1. Bacterial Detection. Bacteria at low concentrations
are hard to detect and usually require long induction times
before further analysis. To detect bacteria at ultralow concen-
trations without time-consuming procedures is advantageous
in clinical diagnosis and environmental monitoring. We devel-
oped a simple strategy that uses vancomycin-conjugated FePt
nanoparticles (FePt@Van conjugates) to capture and detect
pathogens such as vancomycin-resistant enterococci (VRE) and
other Gram-positive bacteria at exceptional low concentra-
tions.6 Figure 2A shows the experimental procedure of bac-
terial detection using the biofunctional magnetic nanoparticles.
The mixing of FePt@Van conjugates with a solution of bacte-
ria results in a sufficient amount of magnetic nanoparticles
binding onto the bacteria because of their strong interactions
(i.e., polyvalent interactions between Van and D-Ala-D-Ala on
the bacterial surface). A small magnet attracts and enriches
these bacteria-nanoparticle composites for the analysis. In the
control experiment, the use of FePt nanoparticles capped with
FIGURE 2. (A) Interaction between FePt@Van nanoparticles and
nonspecific groups (FePt-NH2) fails to capture the bacteria
bacteria. (B) FePt-NH2 nanoparticles as the control. SEM images of
because of the lack of specific molecular recognition (Figure (C) aggregates of S. aureus and FePt@Van nanoparticles and (D)
2B). aggregates of FePt-NH2 nanoparticles. (E) SEM and (F) TEM images
of a VanA bacteria captured by FePt@Van nanoparticles (inset,
Scanning electron microscope (SEM) can easily distinguish
HRTEM of FePt@Van nanoparticles). Reproduced from ref 6.
the bacteria from the aggregates due to their micrometer Copyright 2003 American Chemical Society.
sizes. FePt@Van conjugates capture bacterial strains such as centrate Gram-negative bacteria such as Escherichia coli due
Staphylococcus aureus, S. epidermidis, and coagulase nega- to the partial exposure of the proper receptors on the surface
tive staphylococci (CNS) with ultralow concentrations. Figure of cells.7,8
2C shows the SEM image of “magnetized” S. aureus aggre- The detection limit achieved using FePt@Van magnetic
gate with FePt@Van conjugates. When FePt-NH2 is used, the nanoparticles is comparable to that of assays based on poly-
SEM image (Figure 2D) shows no S. aureus, suggesting that merase chain reaction (PCR), and this protocol is faster than
FePt@Van binds to S. aureus specifically due to the molecu- PCR when the bacterium count is low. In addition, using bio-
lar recognition. Figure 2E shows the SEM image of vancomy- functional magnetic nanoparticles to capture bacteria is par-
cin-resistant bacteria (VanA genotype) captured by the ticularly useful when PCR is inapplicable. The procedure that
FePt@Van nanoparticles. Transmission electron micrograph combines FePt@Van biofunctional magnetic nanoparticles with
(TEM, Figure 2F) clearly confirms that the nanoparticles bind fluorescence dyes can achieve quick, sensitive, and low-cost
on the surface of bacterial cells. The currently achieved detec- detection of bacteria in blood.9 FePt nanoparticles provide the
tion limit can be as low as 4 cfu/mL. The high-sensitivity bac- platform for introducing Van in order to generate multivalent
terial detection using FePt@Van polyvalent nanoparticles may interactions. The fluorescent vancomycin, conjugate of Van
be due to their similar size to antibodies such as IgG. More- and fluorescein amine (Van-FLA), stains the enriched bacteria
over, FePt@Van nanoparticles can also capture and precon- for the quick detection using fluorescence microscopy. This

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Multifunctional Magnetic Nanoparticles Gao et al.
FIGURE 3. Detecting bacteria in blood samples: (i) addition of FePt@Van; (ii) capturing bacteria assisted by a magnet; (iii) addition of Van-
FLA for staining the bacteria; and (iv) magnetically separating stained bacteria from Van-FLA solution. Right, a fluorescent image of stained
bacteria after the capture. Adapted from ref 9. Copyright 2006 Wiley-VCH.
protocol allows the detection of bacteria from the samples
within 2 h and has sensitivity as low as 10 cfu/mL.9 Figure 3
illustrates the straightforward steps for detecting bacteria in a
blood sample: (i) mixing, (ii) separation, (iii) staining, and (iv)
washing. Using the fluorescence microscope, one can easily
observe the captured bacteria (Figure 3).
Although it is possible to detect the species of bacteria by
combining biofunctional magnetic nanoparticles and a spe-
cific antibody, it may have limited accuracy and render high
cost. In order to quickly and accurately detect the type of
strains at low concentration, the integration of magnetic nano-
particles for bacteria accumulation and the PCR method for
DNA analysis may confer some advantages. After all, biofunc-
tional magnetic nanoparticles would help to open a new and
attractive avenue for pathogen detection and disease diagno-
sis applications.
2.1.2. Protein Purification. Purification and effective
manipulation of proteins is important for their studies and
applications in life sciences. Among the existing protocols,
magnetic separation and purification is a convenient method
for selective and reliable capture of specific proteins, genetic
materials, organelles, and cells.10,11 For example, using
dopamine as a robust anchor,12 the successful synthesis of
NTA-terminated magnetic nanoparticles offers a simple and
versatile platform for separating six histidine (6xHis)-tagged
protein.13,14 The high surface-to-volume ratio and the good
dispersity of the nanoparticles also increase the protein bind-
ing capacity. The target proteins cover the surface of the nano-
particles effectively and quickly, thus reducing the overall
unoccupied surface area for nonspecific absorption of pro-
teins and achieving higher specificity than microparticles.
Moreover, the use of NTA-terminated magnetic nanoparticles
eliminates the pretreatment of the cell lysate because of the
high specificity of the nanoparticles (Figure 4A,B). It is worth
mentioning that the NTA-modified magnetic nanoparticles are
reusable without losing efficiency (Figure 4B). Although pro-
tein purification based on 6xHis-tag has been widely used, the
1100 ACCOUNTS OF CHEMICAL RESEARCH 1097-1107 August 2009
toxicity of the metal-NTA complexes15 in this technique
requires caution for in vivo applications. The specificity of the
magnetic nanoparticles exhibited in protein separation sug-
gests that magnetic nanoparticles, as a general and versatile
system, should selectively bind with other biological targets at
low concentrations if proper anchors and ligands are used.
2.1.3. Toxin Decorporation. Magnetic nanoparticles also
play an important role in toxin decorporation,16 a strategy that
reduces and removes toxins from contaminated bodies. For
example, biofunctional magnetite nanoparticles decorated by
bisphosphonate (BP), which coordinates to a uranyl ion (UO22+)
with high affinity, can remove UO22+ efficiently (Figure 4C).
The designed magnetic nanoparticles, Fe3O4-BP, remove
99% and 69% of UO22+ from water and blood, respectively
(Figure 4D).17 Besides being the first example of removal of
radionuclides from a biological fluid by nanoparticles, this
result suggests that functionalized, biocompatible magnetic
nanoparticles can act as useful and effective agents of deco-
rporation for selectively and rapidly removing radioactive tox-
ins in vivo. Although the successful protocol for removal of
UO22+ using Fe3O4-BP nanoparticles provides a potential plat-
form to develop a biocompatible methodology for decorpo-
rating radioactive hazards from human bodies, it is important
for researchers to demonstrate such a possibility in animal
models as the next phase of research.
2.2. Dyes or Drugs. It is also attractive to conjugate mag-
netic nanoparticles with other functional molecules. Because
organic dyes (e.g., Cy5.5, FITC) have widely served as probes
to visualize biological processes and magnetic nanoparticles
can act as MRI contrast agents, the conjugation of magnetic
nanoparticles and dyes will offer the multifunctional nano-
probes combining MRI and optical imaging.18 For example,
the conjugation of Fe3O4 nanoparticles and porphyrin19 may
lead to a bimodal imaging nanoprobe. Moreover, the well-
studied pharmacokinetics and low systemic toxicity of por-
phyrin derivatives have already led to some clinical trials and
usage of porphyrin derivatives in photodynamic therapy (PDT),
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FIGURE 4. Protein purification and toxin decorporation using biofunctional magnetic nanoparticles: (A) NTA-terminated magnetic
nanoparticles selectively binding to histidine-tagged proteins; (B) SDS/PAGE analysis of the fraction of proteins washed off the magnetic
nanoparticles by imidazole solution at 10 (lane 3), 80 (lane 4), and 500 mM (lane 5) and the fractions washed off the reused nanoparticles
using imidazole solution at 10 (lane 6), 20 (lane 7), and 500 mM (lane 8); (C) Fe3O4-BP nanoparticles remove UO22+ from blood; (D) the
amount of UO22+ in blood (I) before and (II) after the removal, and (III) the amount of UO22+ on the magnetic nanoparticles.

so the porphyrin-modified Fe3O4 nanoparticles can act as a
multifunctional nanomedicine that combines PDT anticancer
treatment and noninvasive MR imaging. Although fluores-
cence imaging may be the fastest growing area for in vivo
analysis, limited tissue penetration of the fluorescence restricts
its application in small animal imaging, which requires organic
fluorophores emitting at above 700 nm in the near-infrared
(NIR). However, there are very limited options for available
NIR-emitting organic probes, and the potential cytotoxicity and
stability of organic fluorophores for in vivo experiments should
be of concern.
3. Combination of Magnetic Nanomaterials
and Other Nanocomponents
Nanostructures provide an excellent platform to integrate differ-
ent functional nanocomponents into one single nanoentity to
exhibit multifunctional properties. To assemble different nano-
particles into a single entity as the novel building block itself is
exciting and holds great potential. As briefly discussed in the fol-
lowing sections, based on the magnetic nanoparticles, one can
combine QDs to exhibit magnetic and fluorescent
properties,20-22 sequentially grow metallic nanocomponents23 or
form exotic nanostructures, such as yolk-shell nanoparticles for
the exploration of nanomedicines.24,25
3.1. Quantum Dots. Several useful applications in the
study of subcellular processes of fundamental importance in
biology have highlighted the potential of QDs in nanobiotech-
nology.26 As now well-recognized, QDs have much greater
temporal stability and resistance to photobleaching than flu-
orescent dyes do. Moreover, there are many alternatives in
QDs with NIR emission for in vivo imaging compared with
organic fluorophores. Such unique and attractive properties of
the QDs have inspired the fabrication of hybrid nanostruc-
tures that exhibit both fluorescence and magnetism, such as
Co@CdSe core-shell nanocomposites27 and FePt-ZnS nano-
sponges.28
Based on the FePt nanoparticles and semiconducting chal-
cogenide nanocomponents, systematic studies show that the
reaction conditions control the formation of different hybrid
nanostructures. In one-pot reaction, the sequential growth of
CdX (X ) S or Se) onto FePt nanoparticles under the lower
reaction temperature results in the formation of FePt@CdX
core-shell nanoparticles (Figure 5A,B).21 However, the use of
a higher boiling point solvent gives FePt-CdX heterodimer
nanoparticles (Figure 5C,D).20,21 The formation of het-
erodimers at higher temperature is probably due to the dif-
ference in phase transition temperatures between the FePt and
CdX components. The CdX components may melt at the
higher temperature and induce dewetting from the FePt cores,
resulting in the formation of heterodimeric nanostructures. The
synthesis of these core-shell and heterodimer nanoparticles
is highly reproducible and general. Although the optical prop-
erties of these nanostructures still need to be improved for
achieving higher quantum yields, this rather simple approach

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Multifunctional Magnetic Nanoparticles Gao et al.
FIGURE 5. Formation of core-shell or heterodimer nanostructures
by FePt and CdX (X ) S or Se). The core-shell nanostructures of (A)
FePt@CdS and (B) FePt@CdSe nanoparticles (insets, HRTEM images).
The heterodimeric nanostructures of (C) FePt-CdS and (D)
FePt-CdSe.20,21 Reproduced in part from ref 21. Copyright 2007
American Chemical Society.
opens an avenue for designing and synthesizing sophisticated
and multifunctional nanostructures.
The combination of superparamagnetism and fluorescence at
nanometer scale should help the biological applications of mul-
tifunctional nanomaterials. Although the fluorescence of QDs is
partially quenched by metallic nanoparticles in FePt-CdX hybrid
nanostructures, the replacement of metallic nanoparticles with
metal oxide nanoparticles results in a good quantum yield. For
example, the Fe3O4-CdSe heterodimer nanoparticles show the
emission wavelength peak at 610 nm with quantum yield of
about 38%. The resulting fluorescent magnetic nanoparticles
bear two attractive features with high quality, superparamag-
netism and fluorescence, which allow their intracellular move-
ments to be controlled using magnetic force and to be monitored
using a fluorescent microscope.22 As shown in Figure 6, the con-
focal microscopic imaging of cells reveals the movement of
Fe3O4-CdSe nanoparticles inside the cells upon applying a mag-
netic force. After incubation, HEK293T cells can take up the GSH-
modified Fe3O4-CdSe nanoparticles together with pEGFP-N1
vector. Without the application of magnetic force, Fe3O4-
CdSe@GSH nanoparticles distribute nonspecifically inside the
cells because of their simple surface modification (Figure 6E). The
homogeneous fluorescent spots suggest that the nanoparticles
mainly distribute in the cytosol. After being attracted by a small
magnet, the Fe3O4-CdSe@GSH nanoparticles aggregate on the
side of the cell nearest the magnet (Figure 6F). Because of their
significant magnetic moment, the magnetic nanoparticles drift to
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FIGURE 6. Intracellular manipulation of Fe3O4-CdSe: (A)
distribution of the nanoparticles in a cell without a magnetic field;
(B) movement of the nanoparticles with the magnetic field; (C)
aggregation due to strong magnetic dipolar-dipolar interactions
(FD-D); (D) the aggregates of the nanoparticles inside the cells. The
confocal images of HEK293T cells after being incubated with the
Fe3O4-CdSe@GSH nanoparticles and pEGFP-N1 vector (E) without
and (F) with a magnetic field. Reproduced from ref 22. Copyright
2008 American Chemical Society.
the magnet due to the magnetic field gradient (H). However, the
movement of each nanoparticle is partially hindered inside the
cells due to the high viscosity of the cytosol, resulting in a rather
slow response. When the nanoparticles approach each other, the
magnetic dipolar-dipolar interactions (FD-D) become the domi-
nant forces and drive the nanoparticles much closer to each other
(Figure 6C). After several hours, the interplay of these two forces
leads to the result of round aggregates of nanoparticles inside the
cells.
The successful demonstration of intracellular manipulation
of magnetic nanoparticles will be useful for biological appli-
cations because this strategy may offer a useful tool to inves-
tigate the difference between the apical and basolateral
domains in polarized cells by the asymmetric localization of
magnetic nanoparticles that carry specific ligands. To realize
these promises, the fluorescent magnetic nanoparticles should
have fast response to the magnetic force, which has yet to be
improved.
 Multifunctional Magnetic Nanoparticles Gao et al.

FIGURE 7. Two approaches for synthesis of Fe3O4-M (M ) Ag or Au) heterodimers: (A) Fe3O4-Ag heterodimers form at a liquid-liquid
interface; (B) TEM and (C) HRTEM images of Fe3O4-Ag heterodimers;23 (D) the formation of Fe3O4-Au heterodimers by the decomposition of
Fe(CO)5 on the surface of Au nanoparticles; (E) TEM and (F) HRTEM images of Fe3O4-Au heterodimers.30

3.2. Metallic Nanoparticles. Metallic nanoparticles (e.g.,
Ag, Au, and Pt) have fascinated scientists for several centu-
ries, partly because of the colorful colloidal solutions of metal-
lic nanoparticles. Because of their excellent optical properties,
metallic nanoparticles are especially useful for biomedical
applications, such as optical contrast agents, multimodal sen-
sors combining optical imaging and scattering imaging, and
photothermal therapy.29 The combination of metallic nano-
particles and magnetic nanoparticles likely will lead to new
applications in biomedicine.
One of the simplest and most efficient methods is the
sequential growth of metallic components (e.g., Ag or Au) onto
a “colloidosome” (i.e., the self-assembly of nanoparticles at a
23
liquid-liquid interface) of magnetic nanoparticles. The
metallic nucleation takes place on the exposed surface of the
magnetic nanoparticles and produces the heterodimers of two
distinct nanospheres (Figure 7A). TEM and HRTEM images (Fig-
ure 7B,C) show Fe3O4-Ag heterodimers with relative uniform
size and structure, indicating the method of liquid-liquid inter-
face heterogeneous growth is a general way to make het-
erodimer nanoparticles. Recently, Sun et al. reported another
way to fabricate Fe3O4-Au heterodimers in a homogeneous
organic solvent,30 where the thermal decomposition of
Fe(CO)5 onto the surface of the Au nanoparticles and the fol-
lowing oxidation of intermediate result in uniform Fe3O4-Au
heterodimers (Figure 7D-F).
The heterodimer structure offers particles with two distinct
surfaces. Different kinds of functional molecules can covalently
bind to the specific parts of the heterodimers. For instance,
Fe3O4 can support a specific biomolecule for targeting using
dopamine as a robust anchor.12,13 For the Ag or Au compo-
nent, one can use the well-developed gold-thiol chemistry for
the attachment of thiol-terminated biomolecules.5,31 Together
with their own distinct functionalities, these multifunctional
heterodimers can respond to magnetic forces, show enhanced
resonance absorption and scattering, and bind with specific
receptors. Using epidermal growth factor receptor isoform A
(EGFRA)-conjugated Fe3O4-Au heterodimer nanoparticles, Sun
et al. demonstrated their dual-functional imaging property for
cell tracking.32 This type of heterodimer nanoparticle may
have great potential in multimodal biomedical applications,33
especially for molecular imaging, but a substantive amount of
work need to be done to achieve these promising applications.
3.3. Yolk-Shell Nanostructures. Using magnetic nano-
structures as drug delivery carriers is an active and promis-
ing research subject on biomedical applications.34 The
conventional and straightforward strategy for drug delivery
using magnetic nanoparticles is the use of polymers to coat
magnetic nanoparticles and to encapsulate drugs to form
nanocapsules or micelles,35 which may require complicated
processes and result in modest efficiency. Based on the nano-
scale Kirkendall effect reported by Alivisatos’s group,36 we

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Multifunctional Magnetic Nanoparticles Gao et al.
FIGURE 8. The ultrahigh cytotoxicity of FePt@CoS2 yolk-shell nanoparticles to HeLa cells: (A) TEM image of FePt@CoS2 yolk-shell
nanoparticles; (B) MTT assay results of FePt@CoS2 nanoparticles on HeLa cells; (C) representative TEM image of FePt@CoS2 nanoparticles in
mitochondria of HeLa cells; (D) the possible mechanism accounts for FePt@CoS2 nanoparticles killing HeLa cells. Reproduced from ref 24.
Copyright 2007 American Chemical Society.
developed FePt@CoS2 yolk-shell nanoparticles as a poten-
tial nanodevice for controlled drug release.24 The unique
structure and properties of FePt@CoS2 yolk-shell nanopar-
ticles give an unconventional example of a novel drug deliv-
ery system that uses the magnetic nanomaterials to directly
encapsulate the potential anticancer drug. The premise is that
platinum-containing nanoparticles (e.g., FePt) without any sur-
face coating may act as potential anticancer drugs, like the
well-known anticancer drug, cisplatin. The sequential growth
of CoS2 porous nanoshells by Kirkendall effect36,37 provides
the feasibility to produce the “naked” FePt nanoyolks and the
special interface between outside and inside of the shells (Fig-
ure 8A).
According to MTT assay results (Figure 8B), FePt@CoS2
nanoparticles show an IC50 value lower than that of cisplatin.
TEM images show that there are many nanoparticles in
organelles such as mitochondria, indicating the cellular uptake
of nanoparticles. More interestingly, there are no more
FePt@CoS2 nanoparticles except some hollow nanospheres in
mitochondria bodies (Figure 8C), indicating that the FePt@CoS2
nanoparticles enter into the organelles and the FePt yolks dis-
1104 ACCOUNTS OF CHEMICAL RESEARCH 1097-1107 August 2009
integrate after the cellular uptake. Figure 8D shows the pos-
sible mechanism of cytotoxicity of FePt@CoS2 nanoparticles
against the HeLa cells: After cellular uptake through the
endocytosis pathway, under the acidic environment inside the
secondary lysosomes, FePt yolks are oxidized (probably by O2
in the presence of oxidase inside cells) and destroyed to pro-
duce platinum(II) species because the reactivity of unprotected
iron promotes the disintegration of FePt. The permeability of
CoS2 shells allows these Pt(II) species to diffuse out of shells,
enter into the nucleus and mitochondria, damage the DNA
double helix chains, and lead to the apoptosis of the HeLa
cells, which is similar to the interaction between cisplatin and
DNA. Although this mechanism (involving DNA damage)
requires more concrete evidence and needs further studies,
the cytotoxicity of FePt@CoS2 nanoparticles clearly originates
from the slow, intracellular dissolution of the FePt yolk, which
has been further confirmed by the toxicity assays of CoS2 hol-
low nanoparticles24 and FePt@Fe2O3 yolk-shell nanopar-
ticles.25 The mechanism implies that intracellular drug release
from a magnetic nanostructure can be a new and powerful
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 Multifunctional Magnetic Nanoparticles Gao et al.

FIGURE 9. TEM images and MTT assay results for HeLa cells of FePt@Fe2O3 yolk-shell nanoparticles (A, E), Pt@Fe2O3 yolk-shell
nanoparticles (B, F), FePt@Fe3O4 core-shell nanoparticles (C, G), and γ-Fe2O3 hollow nanoparticles (D, H). T *-weighted
2 MR images of (I)
FePt@Fe2O3 yolk-shell nanoparticles, (J) Pt@Fe2O3 yolk-shell nanoparticles, (K) FePt@Fe3O4 core-shell nanoparticles, and (L) γ-Fe2O3 hollow
nanoparticles at 25, 50, and 100 µg/mL, respectively. Reproduced from ref 25. Copyright 2008 American Chemical Society.

strategy for delivering therapeutics, which deserves further
exploration and development.
Following the encouraging result of the FePt@CoS2
yolk-shell nanoparticles, we further reported the bifunctional
FePt@Fe2O3 yolk-shell nanoparticles combining high cyto-
toxicity and strong MR contrast enhancement.25 Recent devel-
opment in the synthesis of iron oxide hollow nanoparticles38
facilitates the production of FePt@Fe2O3 yolk-shell nanopar-
ticles. Among four types of nanoparticles (Figure 9A-D),
FePt@Fe2O3 yolk-shell nanoparticles have the highest cyto-
toxicity (Figure 9E-H). Likely due to the same possible mech-
anism of cytotoxicity as FePt@CoS2 yolk-shell nanoparticle,
FePt@Fe2O3 yolk-shell nanoparticles show an ultralow IC50
value in terms of Pt. The Pt nanocrystals in Pt@Fe2O3
yolk-shell nanoparticles are stable because of the relative
inertness of Pt noble metal. The FePt nanoparticles in
FePt@Fe3O4 core-shell nanoparticles are very stable because
the biocompatible, compact Fe3O4 shells prevent the oxida-
tive species from reaching the FePt cores.24,25 These results
suggest two essential requirements of this kind of nanostruc-
ture for serving as an effective anticancer drug: (i) the rela-
tively reactive platinum-containing yolks to form platinum(II)
species; (ii) the good permeability of shells to allow the metal
ions (or complexes) to release from the shells. Obviously,
FePt@CoS2 yolk-shell nanoparticles and FePt@Fe2O3
yolk-shell nanoparticles both satisfy these two requirements

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Multifunctional Magnetic Nanoparticles Gao et al.
and may have effective anticancer activity as potential nano-
medicine. Although the in vivo experiments have yet to be
completed, these results suggest that yolk-shell nanoparticles
may evolve to become useful and effective nanodevices for
controllable drug release in cancer treatment.
T *2-weighted MR image results (Figure 9I-L) indicated
γ-Fe2O3 hollow nanoparticles exhibit the strongest MR signal
attenuation effect among those four types of particles.
FePt@Fe2O3 yolk-shell nanoparticles and Pt@Fe2O3
yolk-shell nanoparticles also show strong MR relaxation
enhancement. It is noteworthy that the hollow magnetic nano-
particles exhibit the advantages in MR relaxation enhance-
ment due to their unique geometry and may attract increased
research interest for their MRI applications. The interface
between the iron oxide hollow nanoparticles and the water
phase is much larger than that of solid iron oxide nanopar-
ticles under the same concentrations, which may contribute to
the stronger MR contrast enhancement per unit weight.25
FePt@Fe2O3 yolk-shell nanoparticles offer several distinct
advantages as a nanodevice model for controlled drug deliv-
ery. First, the biocompatibility of Fe2O3 nanoshells ensures the
origin of cytotoxicity from the “naked” FePt yolk. Second, the
Fe2O3 nanoshells allow cancer-targeting moieties to be
anchored on the shell surface, which would significantly
reduce the side effects. Finally, MR relaxation enhancement
effects of Fe2O3 nanoshells may provide a direct means for
measuring the prognosis during the cancer treatments. Given
the capability of surface functionalization of these yolk-shell
nanoparticles using disease-specific molecules, one could
develop yolk-shell multifunctional nanoparticles that target a
specific tissue for delivering the therapeutic agents and mon-
itoring the transformation of the tumor by MRI. Despite the
lack of in vivo testing results, it is likely that the initial verifi-
cation of these novel magnetic nanoparticles as multifunc-
tional nanomedicines will drive their further development for
multimodal biomedical applications.
4. Conclusion and Perspectives
In this Account, we have summarized some recent design and
synthesis of two classes of multifunctional magnetic nanopar-
ticles: molecular-functionalized and nanocomponent-hybrid-
ized. These two approaches would be complementary to other
methods that integrate the different individual nanoparticles
via polymer coating39 or silica encapsulation.40 Biofunctional
magnetic nanoparticles with high selectivity and high sensi-
tivity not only promise biological applications in bacterial
detection and protein purification, but also offer advantages in
1106 ACCOUNTS OF CHEMICAL RESEARCH 1097-1107 August 2009
tumor targeting and multimodal imaging. The hybrid mag-
netic nanoparticles open up a new avenue to multimode and
multipurpose biomedical applications because of their inte-
grated functions. For example, based on the magnetic nano-
particles as MRI contrast agents, one of the most exciting
applications is the multimodal imaging that integrates opti-
cal imaging (e.g., dyes, QDs, and Au nanoparticles) or positron
emission tomography (PET) imaging (e.g., isotopes) with
MRI.41-44
Although there are many exciting potential biomedical
applications of multifunctional magnetic nanoparticles, con-
siderable challenges and issues remain to be resolved. For
example, heterogeneity of nanomaterials remains a major
problem, and it is hard to precisely control the number of
functional molecules on the surface of nanoparticles.
Researchers need to develop better strategies for producing
nanoparticles that have precise composition, uniform surface
modification, and reproducible functionalization. For in vivo
biomedical applications, the purity, dispersity, and stability of
the multifunctional magnetic nanoparticles in a physiological
environment are highly important.45 Therefore, it is neces-
sary to further study and explore multifunctional magnetic
nanoparticles for creating successful nanobiotechnology of
biomedical applications.
This work was partially supported by RGC (Hong Kong), HIA
(HKUST), and Brandeis University.
BIOGRAPHICAL INFORMATION
Jinhao Gao received his B.S. from Nanjing University and Ph.D.
from The Hong Kong University of Science and Technology. He
received the Hong Kong Young Scientist Award (2007). He cur-
rently is a postdoctoral fellow at Stanford University. His research
interests are biomedical applications of multifunctional
nanomaterials.
Hongwei Gu received his B.S. and M.S. in Chemistry from the
Department for Intensive Instruction (1998) and Department of
Chemistry (2001) at Nanjing University, respectively. He obtained
his Ph.D. in 2004 under the supervision of Professor Xu before
two years’ postdoctoral training with Professor Swager at MIT. As
a Chemistry Professor of Soochow University, his current research
focuses on the development of new nanomaterials as catalysts for
organic synthesis.
Bing Xu received his B.S. and M.S. from Nanjing University in
1987 and 1990. He obtained his Ph.D. in 1996 from the Univer-
sity of Pennsylvania. After finishing NIH postdoctoral training at
Harvard University, he started his independent research at The
Hong Kong University of Science and Technology in August
2000. He was the recipient of the DuPont Asian & European
Young Investigator Award (2001). Professor Xu recently joined
Vol. 42, No. 8

the Department of Chemistry, Brandeis University, and his
research focuses on the applications of supramolecular chemis-
try to materials, nanoscience, and biological science.
FOOTNOTES
* To whom correspondence should be addressed. E-mail: bxu@brandeis.edu.
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